16alpha, 17alpha-cyclosulfate and cyclosulfite esters of steroids



im t oetll Patented Dec. 1S, 19%2 wovon LJLLRS to No Drawing. Filed Apr.24, 1.961, No. loll-,h2o @la-hns. fdl. Zoll-23955;

ln Formula I, the 1,2- and/ or 6,7-positions may be saturated or doublebonded so that A4-, A191-, 34'5- and Alf'S-pregnanes are included Withinthe scope ot' this invention.

The symbols in formula have the following meanings: R representshydrogen, l represents ,tt-hydroxy or together R and R are lieto (C=0);R" represents hydrogen, halogen or lower alkyl; X and X each representshy drogen or halogen, but at least one of these two symbols representshydrogen; Y represents hydrogen or lower alkyl, preferably methyl; Zrepresents hydrogen, halogen, hydroxy or the acyloxy radical of ahydrocarbon carboXylic acid of less than 1G carbon atoms and itrepresents either 1 or 2.

The symbols R, X, and Z represent all tout halogens but chlorine anduorine are preferred in this group. The same or different halogens mayappear in `a given compound. Lower alkyl groups represented by Y and R"include straight and branched chain saturate hydrocarbon radicals suchas methyl, ethyl, propyl, isopropyl, butyl, `t-butyl, amyl and the like.

Carboxylic acids from which the acylo-Xy radical Z may be derived toproduce 21-esters include, for example, alkanoic acids, preferably loweralkanoic acids such as acetic, propionic, butyric, hcxanoic acids andthe like, lower allrenoic acids such as acrylic acid, monocyclicaromatic carboxylic acids such as bcnzoic and o-, mand p-toluic acids,cycloallianoic acids sucn as cyclohexanoic acid, cyclcallienoic acidssuch as cyclohexcnoic acid, and monocyclic aryl-lower allanoic acidssuch as phenylacetic and -phenylpropionic acids.

The following loaelh-cyclosulte and welk-cyclosulfate esters of C-ringsubstituted-16a,17xdihydroxypregnenes are compounds of `this invention.They and the Working examples which follow serve to illustrate the classof esters included.

16a-h ydroxyhydro cortisone 16 et, 17 -sultlte,16a-hydroxyhydrocortisone 160:,l7ct-sulte Zl-acetate, 2st-methyl- 16@hydroxyhydrocortisone 16,17a sulte, 9ct nuoro- 16-hydroxyhydrocortisone160:,171 sulte, 12a chloro- 1ooc-hydroxyhydrocortisone 16a,17c sulte,6a. methyl- 1ct-hydroxyhydrocortisone l6ct,i7snlte,Soz-fluoro-locahydroxyhydrocortisone 16a,17c suliite, 6a,9a diiluoro-16a hydroxyhydrocortisone l6ct,l7ct sulfate, 9ct,21 diwherein the 1,2-and/ or 6,7-positions are saturated or double bonded, R, R', R", X, X',and Y have the same meaning as before and Z represents hydrogen, halogenor the acyloxy radical of a hydrocarbon carboxylic acid of less than 10carbon atoms,

with eitherthionyl chloride to produce the cyclic sulte or sulurylchloride to produce the cyclic sulfate. rl`he reaction is carried out inthe presence of an organic nitrogen base such as pyridine, collidine,triethanolamine, quinoline or the like and the product is recovered fromthe solution by conventional procedures. Preferably the compound offormula li is dissolved or suspended in the basic medium and thentreated with an equimolccuiar proportion or excess of the thionylchloride or sulfuryl chloride at a temperature below about C.

An alternate procedure involves the reaction of the starting materialwith tliionyl chloride as described previously, then oxidizing thecyclic sulte to the sulfate with an oxidizing agent e.g. an alkalipermanganate such as as calcium permanganate.

The starting materials of formula I are known substances or are readilyobtained from known compounds. Thus y1f5ix,17l t-dihydroxy compounds maybe obtained from their known 17a-hydroxy analogs by enzymatichydroxylation at the l-position by means of the microorganismStreptomyces roseocltromogenus according to the method described in US.latent No. 2,855,343. Similarly, compounds saturated in the 1,2-positionmay be converted to the corresponding 1,2-unsaturated cornpound by theaction of Bacterium cyclooxydans according to the method described inExample 1 of U.S. Patent No. 2,822,318.

Compounds of Formula H bearing 12a-halo anu/ or 6er-methyl substituentsmay be produced as described in my copending application Serial No.677,205, tiled August 9, 1957, now abandoned, eg. by subjecting thecorresponding 16-desoxy compound bearing a 12er-halo and/ or 6er-methylsubstituent to the oxygenating action of the microorganism Streptomycesroseochromogenus.

The 21-acyloxy-16a,17adiol starting materials for the compounds of thisinvention can be obtained by treating the corresponding16,17c.,2l-triols with an acid anhydride in pyridine and separating theresulting mixture by fractional crystallization. An alternate, morelengthy but more general procedure involves the treatment of the16u.,17a,2l-triol with a ketone (eg. acetone) or the aldehyde in thepresence of an acid catalyst, (eg. perchloric acid) to yield thecorresponding 16al7oa-ketal or acetal which is then treated with an acylchloride (eg. acetyl chloride) or an acid anhydride (eg. aceticanhydride) in a basic organic medium (eg. pyridine) to form thecorresponding 21-acyloxy-16o,i7ketal or acetal. The latter is convertedby hydrolysis with aqueous formic acid, c g. less than about formic acidat a temperature Within the range of about 40 C. to about 100 C., to thedesired 21-acyloxy-16a,l7ot-diol by the procedure described in mycopending application Serial No. 84,989, tiled lanuary 26, 1961.

The 2l-halo16,17odiol starting materials for the compounds of thisinvention are prepared by converting the corresponding 16e,17a,21-triolto its 16ct,l7a-ketal or acetal as described above, treating the latterwith an orgcu'c sulonyl chloride (eg. tosyl chloride or mesyl chloride)to prepare the 21-sultonyloxy derivative which is 21-halogenated bytreatment with an alkali metal halide (eg. potassium biiiuoride, lithiumchloride, lithium bromide and sodium iodide). The latter is converted tothe desired 21-halo-16a,17adiol compound by hydrolizing o `the;,17oc-acetal or ketal grouping with formic acid.

When a 6,7-saturated steroid is used as the starting material and a-dehydro inal product is desired, the latter may be obtained by treatingthe 6,7-saturated-l6a,l7ct cyclic esters of this invention with adehydrogenating agent capable of selectively dehydrogenating thisposition. A suitable dehydrogenating agent is chloranil in ethyl acetateand acetic acid.

When a 21-acyloxy steroid is employed as the starting material and thecorresponding 21-hydroxy steroid is desired as the iinai product, the2l-acyloxy-16,17edihydroxy 16-l7-cyclic ester product is hydrolized, forexample, by treatment with an alkali metal carbonate, such as potassiumcarbonate, to yield the desired free 21- hydroxy iinal product.

Among the starting materials of formula H which may be used to producethe products of formula I are the following:

The 21-esters of 16x-hydroxyhydrocortisones, such as the 2l-acetates of16a-hydroxyhydrocortis one, Zat-methylu-hydroxyhycortisone 9a-1'iuoro-16oa-hydroxyhydrocortisone,

12a-chloro-16a-hydroxyhydrocortisone,

6er-methyl-16-hydroxyhydrocortisone,

6-fluoro-1 6-hydroxyhydrocortisone,

and aa-diiiuoro-16a-hydroxyhydrocortisone.

The 21-esters of hydroxycortisones such as the 21-acetates of16u-hydroxycortisone, 2er-methyl-lot-hydroxycortisone,9ct-fluoro-la-hydroxycortisone, 12a-fluoro-la-hydroxycortisone,Gu-inethyl-lta-hydroxycortisone, a-chloro-la-hydroxycortisone,6a-*luoro-l-hydroxycortisone, and cc-a-difluoro-lot-hydroxycortisone.

The 21-esters of 1-hydroxyprednisolones such as the 21-acetates of1ct-hydroxyprednisolone, 9ot-uoro-16othydroxyprednisolone, 12a chloro16a hydroxyprednisolone, 6-methyl-l6ahydroxyprednisolone,6er-fluorolot-hydroxyprednisolone, orot-diiluoro 16ahydroxyprednisolone.

The 21-esters of 16a-hydroxyprednisones such as the 21-acetates of 16a-hydroxyprednisone,

9a-uoro- 1 Gfx-hydroxyprednisone, 12a-iluoro-16a-hydroxyprednisone,oc-methyl- 1 -hydroxyprednisone, 6ft-chloro-lot-hydroxyprednisone,uct-difluoro-1a-hydroxyprednisone.

The compounds of this invention are physiologically active substanceswhich possess glucocorticoid and antiinammatory activities and hence canbe used in lieu of known glucocorticoids such as hydrocortisone andcortisone in the treatment of rheumatoid arthritis for which purposethey can be administered in the same manner as, for example,hydrocortisone, the dosage being adjusted for the relative potency ofthe particular steroid. They may be administered orally, for example, inthe form of tablets or capsules by incorporating a therapeutic dosagewith a carrier according to conventional practice.

The following examples are presented to more fully illustrate thepresent invention (all temperatures being expressed in degreescentigrade).

EXAMPLE 1 Trczmcz'nolone 16(2,17a-Sztle 21 -Acetae Am, 2.38 mu(e=l.3,600); am?? 2.80 (shoulder), 2.98,

5.75, 5.82, 6.01, @.20 and Analysis.--Calcd for C23H29O7F (436.46): C,63.29; H, 6.69. Found: C, 63.21; H, 6.81.

B. Preparation of triamcinolone 16,17sulfite 2lacetate: To a solution of1 g. of triamcinolone 21-monoacetate in 30 ml. of pyridine is added withstirring at --15, 2 ml. of thionyl chloride. The reaction is allowed toproceed at 15 for 2.5 minutes after which time ice water is added.Chloroform is added to the mixture and the layers are separated. Thechloroform extract is washed with water, l N sulfuric acid, water,dilute sodium bicarbonate solution and again with water, dried oversodium sulfate and the solvent removed in vacuo. The resulting residue(about 1.2 g.) is dissolved in l ml. of chloroform and 50 ml. of benzeneand chromatographed on 20 g. of neutral alumina. Elution of the columnwith 500 ml. of a mixture of one part of chloroform and 5 parts ofbenzene yields about 560 mg. of crystalline material, which afterrecrystallization from acetone-hexane represents analytically puretriamcinolone l6ot,17osuliite 21-acetate possessing the followingproperties: MP. 288-289; [a]D23+l30 (c, 1.12 in chlf.);

Nuiol 5.74., 5.78, aoc, 62s, 8.25 to $5.29,.

AnaZysis.-Calcd for C23H27O8SF (482.50): C, 57.24; H, 5.64; S, 6.65.Found: C, 57.60; H, 5.68; S, 6.32.

6 EXAMPLE 2 Tramcilzolone 16a,17a-Sulfte To a solution of mg. oftriamcinolone 16,17o sulite-Zl-acetate in 10 ml. of methanol is addedunder nitrogen 1 ml. of a 10%' solution potassium carbonate in water.The mixture is allowed to stand at room ternperature for 1/2 hour afterwhich the mixture is acidifed with 0.1 ml. of glacial acetic acid, 2 ml.water are added and the methanol removed in vacuo. The resultingcrystalline precipitate is filtered olf, washed with water and dried toyield the product triamcinolone 16,17otsulte.

EXAMPLE 3 Trzfzmcinolone I 60a] 7a-Sulfate 21 -Acetate To a solution of100 mg. of triamcinolone 21-acetate in 3 ml. of pyridine is added at -15with stirring 0.1 ml. of sulfuryl chloride. The reaction is allowed toproceed at 15 for 15 minutes, after which time ice water is added andthe mixture extracted with chloroform. The chloroform extract is washedwith water, 1 N sulfurie acid, water, dilute sodium bicarbonate, againwith water, dried over sodium sulfate and the chloroform removed invacuo. The resulting residue (about 200 mg.) after recrystallizationfrom 95% alcohol furnishes 100 mg. of pure triamcinolone 16a,17asulfate21-acetate possessing the following properties: MP. 232 (dec.);[o]D23-}-105 (c, .30 in ch1f.);

my 2.99, 5.72, 5.77, ser, 6.26, @.25 and 8.27,.

Analysz's.-Calcd for C23H27O9FS (498.50): C, 55.40; H, 5.45; F, 6.44.Found: C, 55.38; H, 6.45; F, 6.25.

EXAMPLE 4 6-Dehydrotriamcitolone Z6oL-17ot-Sulte 21--Aceate A solutionof 250 mg. of the product of Example 1B and 1.250 g. of recrystallizedchloranil in 15 rnl. of tert. butanol is heated under reflux for threehours. The mixture is cooled, iiltered, the filtrate poured into waterand the layers separated. After an additional extraction of the aqueouslayer with ethyl acetate, the ethyl acetate extract is washed with a 1 Nsodium hydroxide solution until the aqueous layer becomes colorless. Thewashed extract is dried over sodium sulfate and evaporated to dryness invacuo leaving as residue the product 6-dehydrotriamcinolone1604,17ot-sullite 2l-acetate.

EXAM PLE 5 To a solution of 1 g. of triamcinolone acetonide in 10 ml. ofanhydrous pyridine is added at 1 ml. or" methanesulfonyl chloride. Aftertwo hours at 0 ice water is added and the precipitated rnesylate isremoved by filtration. The precipitate is washed thoroughly with waterand dried in vacuo, then recrystallized from acetone-hexane. Thetriamcinolone acetonide 21-mesylate melts at about 24S-250 (dec.) or286-287 (dec.) (polymorphic forms).

A mixture containing 1 g. of triamcinolone acetonide 21-mesylate, 1 g.of potassium fluoride and 25 ml. of ethylene glycol is retluxed for 19hours. The dark solution is poured into ice water and extracted withchloroform. The chloroform extract is washed with water, dried oversodium sulfate and evaporated to dryness in vacuo. The residue,16a,l7e-isopropylidene-9a, 21-dirluoro A1A pregnadiene -11,16m,17a-triol-3,20 diene, is recrystaliized from acetone with the aid cicharcoal and melts at about 310.

The above product is deacetonated by treatment with formic acid by theprocedure of Example 7 of my coscadere pending application Serial No.84,989, tiled lanuary 26, 1961, i.e. by heating with 88% ormic acid,hydrolyzing with aqueous potassium carbonate and neutralizing withglacial acetic acid, to obtain 9x,2l-difluoronlpregnadiene-1l,l6a,l7ct-triol-3,20dione.

Treatment of the above products with thionyl chloride and pyridine atfor 2.5 minutes in accordance with the procedure of Example 1B yieldsthe iinal product 9oc,2l-difluoro-Aly-pregnadiene l1,16,l7a trici-3,20-dione 16a,l7sulte.

EXAMPLE 6 EXAMPLE 7 oc-Fluorotramcinolone 160:,17ca-Sute .QI-PropiormzeA. Preparation of 6a-lluorotriamcinol0ne 21-propionate: A solution or"200 mg. of 6ct-uorotriamcinolone acetonide in 1 rnl. of pyridine and 0.5rnl. of propionic anhydride is allowed to remain at room temperature fortwenty hours. After removal or" the reagents in vacuo the residue16oc,17cacetonide of 6oa-uorotriamcinolone 21-propionate is deacetonatedwith 60% formic acid at 100 according to the procedure described inExample 5, thereby yielding a-lluorotriamcinolone 2l-propionate.

B. Preparation of a-fluorotriamcinolone l6a.,17usul tite 21-propionate:To a solution of 1 g. of the product of part A in 3G ml. of pyridine isadded, with stirring, at 15", 2 ml. of thionyl chloride. is treated asin Example 1B to yield the product 6a* iluorotriamcinolone16ot,17ast1liite 21-propionate.

EXAMPLE 8 6-Flu0r0triamcinolone 16st,] 7er-Sulfate 21-Propionate To asolution of 100 mg. of 6a-tluorotriamcinolone- 21-propionate in 3 ml. ofpyridine is added at 15 with stirring 0.1 ml. of sulfuryl chloride. Thereaction is allowed to proceed at 15 with stirring for 15 minutes, afterwhich time ice water is added and the mixture extracted with chloroform.The extract is Washed with water, 1 N sulfuric acid, water, dilutesodium bicarbonate, again with Water, dried over sodium sulfate and thechloroform removed lin vacuo. The residue is recrystallized from 95%alcohol to obtain the product, 6er-{lucrotriamcinolone l6,17asulfate21-propionate.

EXAMPLE 9 2oc-Mehyltramlcino[011e 16%] 7a-SLilfite 21 -Acetate 200 mg.of 2-methyl-triamcinolone 21-acetate (prepared by sequentiallyacetonating, acetylating and deacetonating L-methyl triamcinolone inaccordance with the procedure of Example 7A) is dissolved in 3 ml. ofpyridine and treated at 15 With stirring with 2 ml. of thionyl chloridein accordance with the procedure of Example 1B thereby yielding theproduct 2emethyitri anicinolone-le, l7a-sultte-21acetate- The reactionmixture zo U EXAMPLE 10 2 ot-M ethylzrifzmci 11.02011@ l 6 @al7er-Sulfate 21 -A cetae To a solution of l g. of the product of Example9 in 25 ml. of glacial acetic acid is added with stirring and cooling to10-15", a filtered solution of 3 g. of calcium permanganate in 6 ml. ofwater. When the reaction is completed as indicated by the permanentcolor of the solution, the mixture is poured into ice Water andextracted with chloroform. The chloroform extract is washed with water,sodium bicarbonate (the last washes containing some sodium sulfate toremove excess permanganate) and water, and the chloroform extract driedover sodium sulfate. The solvent is removed in vacuo leaving as residuethe product Zot-methyltriamcinolone 16a.l7asulfate 21-acetate.

EXAMPLE 1 l Za-methyl-l6-hydroxyhydrocortisone 21-acetate (prepared bysequentially acetonating, acylating and deacetonatingZa-methyld6er-hydroxyhydrocortisone in accordance with the procedure inExample 7A) is treated in a solution or" pyridine With thionyl chlorideas described in Example 1B thereby yielding the product 2er-methyl-1ct-hydroxyhydrocortisone 16a,l7asulte 21-acetate.

EXAMPLE 12 Zot-M ethyl-1 6 a-H ydroxyhydrocortz'sone ]6x,17-Sulfate21-Acelate Following the procedure of Example 3,20L-methylla-hydroxycortisone 2l-acetate in pyridine is treated withsulfuryl chloride. The reaction yields the product2ci-methyl1a-hydroxycortisone 1605,17a-sulfate 2lacetate.

EXAMPLE 13 The product of the preceding example is treated withrecrystallized chloranil in accordance with the procedure of Example 4yielding the product 2a-methyl-6-dehydro- 1a-hydroxyhydrocortisone16ot,17sulfate Zlaacetate.

EXAMPLE 14 Substitution of l g. of12a-chloro-ldd-hydroxycortisone-Zl-acetate (prepared by sequentiallytreating 12mchloro-la-hydroxycortisone with acetone and perchloric acid,acetic anhydride and pyridine and 60% formic acid in accordance with theprocedure of Example 7A) is treated with thionyl chloride in accordancewith the procedure of Example 1B thereby yielding the product12achloro-lfx-hydroxycortisone 16a,17asuliite 21-acetate.

T he corresponding free 21-ol is obtained by hydrolysis of the21-acetate lwith a solution of potassium carbonate.

EXAMPLE 15 12a-Chiara] 6 a-H ydroxycorfisone l 6 1,1 7 cc-Sul fate 2l -Aceta'te To a solution of mg. of l2achloro-16ochydroxy cortisone2l-acetate in 3 ml. of pyridine is added at 15 with stirring 0.1 ml. ofsuifuryl chloride. The reaction is allowed to proceed at 15 for 15minutes after which time ice water is added and the mixture extractedwith chloroform. The chloroform extract is washed with water, 1 Nsulfuric acid, Water, dilute sodium bicarbonate, again with water, driedover sodium sulfate and the chloro- 9 form removed in vacuo. Theresulting residue after recrystallization from 95% alcohol yields theproduct, 12achloro-la-hydroxycortisone 160:,17a-sulfate ZI-acetate.

EXAMPLE 16 X1-I2a-Chloro-I-Hydroxycortsone 16a,1 7a-Sulfate 21 -Acetateand the 1,2- and 6,7-unsaturates thereof, wherein R represents hydrogen,R represents )Sl-hydroxy and together R and R' are keto; R represents amember of the group consisting of hydrogen, halogen and lower alkyl; Xand X each represents a member of the group consisting of hydrogen andhalogen, at least one representing hydrogen; Y represents a member ofthe group consisting of hydrogen and lower alkyl; Z represents a memberof the group consisting of hydrogen, halogen, hydroxy and the acyloxyradical of a hydrocarbon carboxylic acid of less than 10 carbon atoms;and n represents an integer from l to 2, said halogens being selectedfrom the group consisting of chlorine and fluorine.

2. A compound of the formula CHgO acyl wherein acyl is the acyl radicalof a hydrocarbon carboxylic acid of less than 10 carbon atoms and halois selected from the group consisting of chlorine and uorine.

3. A compound of the formula wherein acyl is the acyl radical of ahydrocarbon carboxylic acid of less than 10 carbon atoms and halo isselected from the group consisting of chlorine and uorine.

4. A compound of the formula C H2O acyl wherein acyl is the acyl radicalof a hydrocarbon carboxylic acid of less than 10 carbon atoms and halois selected from the group consisting of chlorine and uorine.

5. A compound of the formula wherein "acy1 is the acyl radical of ahydrocarbon carboxylic acid of less than 10 carbon atoms and halo isselected from the group consisting of chlorine and uorine.

6. T riamcinolone 16,l7asulite 21-acetate.

7. Triamcinolone :,17a-sulfate 21aceta'te.

10. a-uorotriamcinolone-l6a,l7asullite -21 -propionate.

References Cited in the le of this patent Fieser et al.: Steroids(1959), page 945, Reinhold Publishing Corporation, New York.

1. A COMPOUND SELECTED FROM THE GROUP CONSISTING OF STERIODS OF THEFORMULA